Recombinant HIL-24 Combined With Chemotherapeutics Inhibits Proliferation Of MCF-7 Cells And Mechanism Research

Objective:(1)To investigate the inhibitory effect of rhIL-24,ADM and Que separately,as well as of rhIL-24 in combination with ADM or Que on MCF-7 breast cancer cell growth and apoptosis in vitro;(2)To investigate the effect of rhIL-24 in combination with ADM or Que separately on MCF-7 breast cancer cell proliferation and apoptosis following autophagy inhibition;(3)To investigate the effect of rhIL-24,ADM and Que separately,as well as of rhIL-24 in combination with ADM or Que on Bcl-2 and Beclin-1 expression in breast cancer cell MCF-7.Methods:(1)After treating with rhIL-24,ADM and Que separately,the changes of cellular morphology,proliferation and apoptosis were analyzed every hour by using Essen Incucyte ZOOM;(2)After treating with rhIL-24 in combination with ADM or Que,the changes of cellular morphology,proliferation and apoptosis were analyzed every hour by using Essen Incucyte ZOOM;(3)The changes of morphology,proliferation and apoptosis were analyzed every hour by using Essen Incucyte ZOOM in MCF-7 cells grouping according to the above(method 1 and 2)after pretreatment with 3-MA;(4)After treating with rhIL-24,ADM and Que separately,as well as of rhIL-24 in combination with ADM or Que on MCF-7 breast cancer cell for 48 hours,the expression levels of Bcl-2 and Beclin-1 were detected by Western Blot.Results:(1)The result of single treatment effect: rhIL-24 was not significantly inhibited MCF-7 cells proliferation;ADM shows significant inhibitory effect on MCF-7 cells,among the experimental group,2.0 ?g/ml ADM group displayed strongest inhibitory effect,which inhibition rate up to 48.24% in 48 hours and in concentration dependence;Que shows significant inhibitory effect on MCF-7 cells,among the experimental group,200 ?M Que group displayed strongest inhibitory effect,which inhibition rate up to 33.53% in 48 hours,but not in a time and concentration dependence.(2)The result of combined treatment effect: MCF-7 cells inhibition rate up to 62.6% after 160 ng/ml rhIL-24 in combination with 1.5 ?g/ml ADM treatment in 24 hours;MCF-7 cells inhibition rate up to 35.35% after 160 ng/ml rhIL-24 in combination with 100?M Que treatment in 24 hours.The result above suggesting that rhIL-24 could enhance ADM or Que inhibitory effect on MCF-7 cells.rhIL-24 in combination with ADM or Que enhanced MCF-7 cells apoptosis rate,suggesting that rhIL-24 could enhance ADM or Que apoptotic effect on MCF-7 cells.(3)The result of autophagic inhibition effect: combined treatment with rhIL-24 and ADM reduced the apoptosis of MCF-7 cells after 3-MA pretreatment,suggesting that autophagy could promote apoptosis of MCF-7 cells.(4)The result of Western Blot: combined treatment with rhIL-24 and ADM reduced the expression of Bcl-2 and Beclin-1,while combined treatment with rhIL-24 and Que reduced Bcl-2 level,but enhanced Beclin-1 level.Conclusions:(1)Single treatment of rhIL-24 was not significantly inhibited MCF-7 cells proliferation;(2)rhIL-24 enhanced the antitumoractivities of ADM or Que by promoting cell apoptosis;(3)Autophagy could be one of the mechanism inducing MCF-7 cell apoptosis;(4)By means of combined treatment with rhIL-24,the effect of reducing expression of Bcl-2 by ADM or Que were enhanced,which could be one of the sensitizing mechanism of rhIL-24.